Sulfonamides preparation



i atented June 26 195i i tries SULFONAMIDES PREPARATION Raymond Alfred Delor and Harold G. Petering, Kalamazoo, Mich, assignors to The Upjohn Company, Kalamazoo, Mich., a corporation of Michigan No Drawing. Application August 19, 1948,

' Serial N0. 45,198

2 Claims.

This invention relates to a pharmaceutical preparation containing, as active ingredients, an absorbable sulfonamide in combination with folic acid.

It is well known that the administration of sulfonamides causes extensive changes in the blood picture. These changes, such as granulocytopenia and anemia, as well as others, have been grouped together under the general term of blood dyscrasia. The appearance of blood dyscrasia following the administration of sulfonamide drugs constitutes one of the most important undesirable side effects of such therapy, making it necessary to maintain careful clinical observation of the patients blood. The blood changes may even be of such an extensive and severe nature as to demand cessation of sulfonamide therapy before such is desirable. After an abnormal blood picture has once appeared, it is extremely difiicult to restore it again to normal. This is particularly true when the patient is suffering from a serious disorder other than blood dyscrasia, the correction of which may be very difficult without the use of sulfonamides. It is also believed that the presence of granulocytes in the blood cells is essential for the destruction of infecting bacteria, as the sulfonamides are considered merely to prevent multiplication of, and not actually to kill, those organisms already present, and the reduction in number of these granulocytes by sulfonamide therapy presents a further complication in effecting restoration of the patients health.

Curative measures of any sort are most undesirable as the incidence of total or acute incapacitation upon onset of the blood dyscrasias is very high, and only a limited percent of the patients treated after symptoms of blood dyscrasia have once appeared are capable of complete or ready recovery even over a lengthy period of time. An increased mortality or prolonged incapacity in such cases appears to be unavoidable. It is thus not only impractical but also mortally dangerous to attempt corrective measures for the sulfonamide-produced blood dyscrasia, e. g., administration of corrective agents after soluble sulfonamide blood eiiects have set in. It is further true that such corrections as can be made are only possible upon discontinuation of the sulfonamide therapy, thus leaving the patient in a particularly helpless position if his infection has not yet been corrected and sulfonamide treatment is vitally in order, or susceptible to a further onslaught of the infection if incomplete recovery has been made and recuperative sulfonamide therapy is advis- 2 able. Obviously, corrective therapy is not a satisfactory procedure.

The sulfonamide drugs can be divided into those whose therapeutic effect is not due to their absorption from the stomach or intestines, acting directly upon the bacterial flora of the intestine, and those whose therapeutic effectiveness is dependent upon an efiective concentration in the blood, namely, the absorbable or soluble sulfonamides. It is known that insoluble sulfonamides which are not absorbed from the intestine destroy the normal bacterial flora of the intestine, and that, by such destruction, the synthesis of certain substances needed by the body is prevented. Among the substances, the synthesis of which may thus be prevented, are certain members of the vitamin B complex, in particular folic acid and its conjugates. However, the soluble sulfonamides, the therapeutic effectiveness of which is due to a high blood level, contrastingly do not reduce the bacterial flora of the intestine to an extent such as to interfere with body synthesis, and

dyscrasia appearing upon soluble sulfonamide therapy would not appear to be due to inhibition of folic acid production by the body.

Therefore, the curative administration of folic acid after soluble sulfonamide therapy in an attempt to eliminate the blood dyscrasia effects of the sulfonamide treatment is not only unsuggested, since soluble sulfonamides do not decrease the amount of folic acid synthesized by the body, but would also be inadequate even if some beneficial result were obtained.

The use of preventive folic .acid therapy has appeared impossible up to the present time, for it isknown that folic acid is substituted para-aminobenzoic acid which, upon subjection to physiologic action of the body, is believed to dismutate into its moieties, including the para-aminobenzoic acid moiety. Para-aminobenzoic acid has the capacity of preventing blood dyscrasia when administered together with soluble sulfonamides, but unfortunately renders the sulfonamide entirely or very nearly completely ineffective bacteriostatically or bactericidally, and hence the combination of soluble sulfonamides and paraaminobenzoic acid, while eliminating the blood dyscrasia effect of the sulfonamide treatment, also eliminates completely any beneficial effects of the sulfonamide. No reason is apparent why the same eiiect should not be produced by a substituted para-aminobenzoic acid, in which case the combination wouldalso be without therapeutic utility.

It was totally unexpected to us that the combination of folic acid, a substituted para-aminobenzoic acid, and a soluble sulfonamide should not only prevent the blood dyscrasia but also allow unaltered effectiveness of the soluble sulfonamide.

It is therefore an object of the present invention to provide a preparation which prevents the appearance of blood dyscrasia, in animals and man, due to the administration of soluble sulfonamides. It is an additional objectof this invention to provide a soluble sulfonamide preparation which prevents the appearance of blood dyscrasia without interfering with the therapeutic efiectiveness of the sulfonamide. Other objects of the invention will become apparent hereinafter.

It has now been found that the simultaneous administration of suitable dosage of folic acid together with a soluble sulfonarnide surprisingly results in no decrease in the therapeutic efiieiency of the sulfonarnide in the blood stream but does prevent the appearance of an abnormal blood pice ture. The simultaneous administration of folic acid with soluble sulfonamides moreover counteracts to a great extent the retardation of the growth rate of young animals produced by sulfonamides alone. Obviously such finding has great importance.

According to the present invention, the folic acid which is administered simultaneously with the soluble sulfonamide drug may be that contained in a naturally occurring folic-acid-containing material such as aqueous liver extract or dried yeast, or may be synthetic or purified material, or a physiologically active derivative of folic acid. A preferred embodiment of this invention contemplates employment of a highly purified or synthetic folic acid or a physiologically active derivative thereof, either wholly or at least in part.

It is of importance that at least 50 gammas (0.05 milligram) of folic acid or its equivalent as a derivative per gram of sulfonamides be administered simultaneously with the sulfonamide. More may be administered, the maximum being that quantity which, in combination with the total daily dose of the soluble sulfonamide, will furnish for the specie of animal to be treated, the daily requirement for folic acid of that particular species of animal under sulfonamid therapy. A representative formula which may be used in the practice of the present invention consists of Mg. Sulfadiazine 167 Sulfamerazine 16'7 Dried aqueous liver extract 167 Folic acid 0.25

The dried aqueous liver represents three grams of fresh liver. In addition such excipients, binders, and lubricants may be used as to enable this mixture to be made into a compressed tablet. Alternatively the formula may be used as a powder, although such is not a preferred form of administration. The composition may also be ad ministered in the form of a syrup. These products are useful for oral administration in the treatment of animals, such as dogs and cats, as well as in the treatment of man.

The following examples are illustrative only and are not to be construed as limiting:

Example 1 Twenty-eight-day old weanling white rats of random sex, 45-50 grams in weight, were placed on a basal purified diet composed of 72 percent White cells, 9,700 per cubic millimeter Granulocytes, 1,130 per cubic millimeter Red cells, 8,000,000 per cubic millimeter Hemoglobin, 14.2 grams per 100 milliliter Hematocrit 42.0 volume percent Eramplc 2 One percent of sulfathiazole was added to the basal diet of Example 1. At the end of 36 days, seven of the original ten experimental animals had survived, those surviving having gained an average of 18 grams per animal. An examination of the blood of these animals showed on the average:

White cells 7,500 pm. cubic millimeter Granulocytes, per cubic millimeter Red cells, 7,500,000 per cubic millimeter Hemoglobin, 11.9 grams per milliliter Hematocrit 41.0 volume percent Example 2A One hundred gammas of folic acid per gram of sulfonamide was added to the basal diet containing one percent sulfathiazole used in Example 2, and this supplemental diet fed to ten animals. At the end of 36 days seven of the animals had survived, those surviving having gained on the average of 30 grams each. The blood picture was as follows:

White cells, 14,300 per cubic millimeter Granulocytes, 1,860 per cubic millimeter Red cells, 8,500,000 per cubic millimeter Hemoglobin, 13.3 grams per 100 milliliter Hematccrit, 44.0 volume percent Example 2B A group of ten animals were fed the basal diet plus one percent sulfathiazole used in Example 2 to which had been added 2.0 grams of liver powder, 1 :20 (containing 100 gammas offolic acid) per gram of sulfathiazole. At the end of 36 days, eight of the ten animals had survived, those surviving having gained on the average 40 grams. An examination of the biood of the surviving animals showed on the average:

White cells, 15,200 per cubic millimeter Granulocytes, 1,220 per cubic millimeter Red cells, 7,600,000 per cubic millimeter Hemoglobin, 12.6 grams per 100 milliliter Hematocrit, 43.0 volume percent Example 3 In a manner similar to Example 2, a diet containing one percent sulfanilamide was fed to a group of ten white rats. At the end of 56 days, nine of the ten animals had survived, gaining an 5 6 average of 45 grams each. The average blood Red cells, 7,600,000 per cubic millimeter picture was: Hemoglobin, 9.8 grams per 100 milliliter He tocrt, 2. v lume ercent White cells, 7,100 per cubic millimeter 1 3 o p Granulocytes, 430 per cubic millimeter 5 Example 4B Red 0611336900900 per Cubic To the diet of Example 4 there was added 100 Hemoglobin, grams per 100 mllhhter gammas of folic acid per gram of sulfadiazine as Hematocnt, Volume percent 1.0 gram of dried 1:20 liver extract. Ten of Example 3A ten animals survived the 56 days of feeding, gainb ing an average of 58 grams each. The average When 50 gammas of folic acid per gram of b1 01 icture wa sulfanilamide was added to the diet of Example White cells, 600 per cubic millimeter all animals Survived gained an allemge Granulocytes,1,400 per cubic millimeter of 60 grams each. The average blood picture I Red cells, 7,900,000 per cubid millimeter was: 15 Hemoglobin, 10.2 grams per 100 milliliter White cells, 17,400 per cubic millimeter crit, 40.0 volume percent Granulocytes, 2,440 per cubic millimeter Example 40 Red cells, 7,400,000 per cubic millimeter Hemoglobin, 12.2 grams per 100 milliliter Hematocrit, 40.0 volume percent To the diet of Example 4 there was added 100 ganimas of folic acid per gram of sulfadiazine as represented by 2.0 grams of dried yeast extract Emmple 33 per 100 grams of diet. Nine of ten animals survived the 56 days of feeding, gaining on the aversulfanilamide, contained in 1000 milligrams of age of 35 grams each The average blood 1:20 liver powder, was added to the diet of Exture was: ample 3, at the end of 56 days all animals had White cells, 3001 cubic l ter When 50 gammas of folic acid per gram of survived, gaining an average of 60 grams each. Granlllocytes, 1,309Def011bic millimeter The average blood picture was: Red cells, 7,200,000 per cubic millimeter Hemoglobin, 11.8 rams er 100 milliliter White cells, 18,500 per cubic millimeter Hematocrit 420 folumelgercent Granulocytes, 2,590 per cubic millimeter Red cells, 7,500,000 per cubic millimeter pl 5 Hemoglobin, grams Per 100 mlnlhter To a basic diet as in Example 1, there was added Hematocrlt, Volume Percent in separate experiments (1) 0.5 per cent sul- Ewample 4 fadiazine and 0.5 percent sulfamerazine, and (2) the above quantities of sulfonamides and 750 To the basic diet of Example 1 there was added gammas of folic acid per 100 grams of diet. At

sulfadiazine one percent by weight, At the end the end of 56 days the following results were of 56 days, ten of ten animals survived, having noted:

White Hemato- Hemo- Granule- Red Cells Cells 3 cnt Vol. globm/lOO cytes er peflmm) 5g Perbent cc. (mmga Basal Diet only 8,390 000 12,870 44. 1 Diet plus 0.5% Sulfadiazine and 0.5% Sulfameraziue 6, 060: 000 7, 740 31.; 3 0 Diet plus Sulionamicles plus Folic Acid 750 gammas per 100 grams diet 7,580,000 14, 820 39.0 12.1 7 1,820

ain an avera e Wei ht f 20 rams each. The g ed g g Example 6 average blood picture was.

White cells, 8,300 per cubic millimeter With a basic diet as in Example 1, the follow- Granulocytes, 130 per cubic millimeter mg supplements were added. After 56 days of Red cells, 5,900,000 per cubic millimeter feeding the red cell count, white cell count, Hemoglobin, 8.1 grams per 100 milliliter hematocrit value, hemoglobin and granulocytes Hematocrit, 32.0 volume percent were determined to be as follows:

White Hcmnto- Hcnio- Granulo- Red Cells Cells Cll',V01. globin/IOO cytes er per(mm')3 35 Per Cent cc. (mung- Basal Diet Only 8, 940, 000 12,460 40.1 13.7 g. 1,170 Basal Diet plus 0.33 per cent sulfathiazole, 0.33 per cent sulfadiazine, 0.33 per cent sulfamerazine 5, 790, 000 T, 040 32. 4 9. 45 310 Diet with the following added per 100 grams of diet: 0.33%

sulfathiazole, 0.33% sulfadiazine, 0.33% sulfamerazine, 1,000 gammas folic acid 7, 900, 000 15, 700 39. 6 12. 2 1,600

Example 4A Various modifications may be made in the present invention without departing from the spirit or scope thereof, and it is to be understood that we limit ourselves only as defined in the appended claims.

To the diet of Example 4 was added 50 gammas of folic acid per gram of sulfadiazine. Nine of ten animals survived after 56 days of feeding, having gained an average of 40 grams each. The

1 We claim: t blood plcture 1. A compressed tablet comprising about 167 Wh1tece1 s, 2 p r cubw mllhmeter milligrams of sulfadiazine, about 167 milligrams Granulocytes, 1,060 per cubic millimeter of sulfamerazine, about 167 milligrams of dried aqueous liver extract, and about 0.25 milligram of folic acid, said tablet upon oral administration producing efiective blood levels of sulfadiazine and sulfamerazine without substantial injurious effect on blood cells and hemoglobin.

2. A therapeutic composition comprising about one part of suliadiazine, about one part of sulfamerazine, about one part of dried aqueous liver extract, and about .9015 part of folic acid. said composition upon oral administration producing eifective blood levels of sulfadiazine and sulfa-- merazine without substantial injurious effect on blood cells and hemoglobin.

RAYMOND ALFRED DELOR. HAROLD G. PETE-RING.

REFERENCES CITED The following references are of record in the file of this patent:

Proc. Soc. Exptl. Biol. and Med. 49 (1942), pages 618-621.

Manufacturing Chemist, February 1947, page 85.

Proc. Soc. Exptl. Biol. &: Med, April 1947, pages J. Biol. Chem., October 1946, pages 439-440.

A ch ve o Bio h m r Ap i 94 a e 9 to 14.

Science, July 2, 1943, pages 20 to 22. 

1. A COMPRESSED TABLET COMPRISING ABOUT 167 MILLIGRAMS OF SULFADIAZINE, ABOUT 167 MILLIGRAMS OF SULFAMERAZINE, ABOUT 167 MILLIGRAMS OF DRIED AQUEOUS LIVER EXTRACT, AND ABOUT 0.25 MILLIGRAM OF FOLIC ACID, SAID TABLE UPON ORAL ADMINISTRATION PRODUCING EFFECTIVE BLOOD LEVELS OF SULFADIAZINE EFFECT ON BLOOD CELLS AND HEMOGLOBIN. 